Annelated dihydropyridines and the use thereof for preparing pharmaceutical preparations

ABSTRACT

A compound of formula I ##STR1## wherein A denotes a benzo, indolo or thieno group; 
     R 1  denotes thienyl or the group ##STR2## wherein R 7 , R 8  and R 9  independently of one another may represent methyl, ethyl, propyl, phenyl or benzyl, while not more than 2 of the substituents can simultaneously represent phenyl or benzyl; 
     R 2 , m, R 3  and R 4  are defined as in the specification, as well as pharmaceutical preparations containing this compound and the new pharmaceutical uses thereof.

This is a continuation of application Ser. No. 08/360,524, filed Dec.21, 1994, now U.S. Pat. No. 5,607,943.

The invention relates to new annelated dihydropyridinoacetic acidderivatives, processes for preparing them and pharmaceuticalcompositions containing these compounds.

Dihydroisoquinolines are known from EP-A 37 934. The compounds specifiedtherein are cardiotonically active and have the effects of increasingcontractility and influencing blood pressure. They have been proposedfor improving blood circulation through the tissues and for improvingthe oxygen supply to the tissues. These possible uses are based on thevascular activity of the compounds. EP-A 251 194 and EP-A 288 048describe how carbocyclically and heterocyclically annelateddihydropyridines have a cardioprotective or cerebroprotective activityand constitute an entirely new type of Ca-antagonistic compounds. WO92/11010 describes the use of such compounds for cerebroprotectiveagents, for treating chronic inflammatory processes and for inhibitingblood clotting and blood platelet aggregation.

The present invention relates to new carbocyclically andheterocyclically annelated dihydropyridines and the pharmaceutical useof these compounds. The new compounds have valuable therapeuticallyuseful properties. They may be used as cardioprotective agents, ascerebroprotective agents (particularly for treating patients who havesuffered a stroke or are in danger of suffering a stroke) and as agentsfor treating chronically inflammatory processes (e.g. bronchial asthmaand arthritis). These compounds may also be used as agents with anantiproliferative effect and as agents for treating ulcerative colitisand Crohn's disease.

The invention relates to compounds of general formula I ##STR3## whereinA denotes a benzo, indolo or thieno group;

wherein, if A is benzo, m is 2 or 3 (preferably 2, whilst the two R² sare in positions 6 and 7) and the substituents R² independently of eachother denote hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I),(C₁₋₄)alkyl, methanesulphonyloxy or methanesulphonamido, or two adjacentsubstituents R² may together represent --O--CH₂ --O-- or --O--CH₂ --CH₂--O--; and if A is indolo or thieno, m is zero;

R¹ denotes thienyl or the group ##STR4## wherein R⁷, R⁸ and R⁹independently of one another may represent methyl, ethyl, propyl, phenylor benzyl, whilst not more than 2 of the substituents can simultaneouslyrepresent phenyl or benzyl;

R³ and R⁴ independently of each other have one of the followingmeanings:

(a) hydrogen,

(b) branched or unbranched C₃₋₆ -alkenyl,

(c) branched or unbranched C₃₋₆ -alkynyl, or

(d) branched or unbranched C₁₋₁₂ -alkyl, wherein the alkyl may besubstituted by

hydroxy,

(C₁₋₄)alkoxy,

di(C₁₋₄)alkylamino,

furyl,

pyridyl,

pyrrolidinyl, N-methylpyrrolidinyl,

morpholino,

indolyl,

nitrilo,

thienyl,

adamantyl,

cyclohexyl,

phenoxy,

naphthyloxy or phenyl, whilst this phenyl or the phenyl contained in thephenoxy group may be mono-, di- or trisubstituted by hydroxy,(C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), CF₃, N₃, CN,(C₁₋₄)alkyl, adamantyl, --SO₂ NH₂, --NHCOCH₃, --NHSO₂ CH₃ or CH₃ SO₂ O--or by the bridge --O--CH₂ --O--;! or by two unsubstituted phenyl groups;

or R³ represents hydrogen and R⁴ represents cyclohexyl, phenyl (whilstthis phenyl may be mono-, di- or trisubstituted by hydroxy,(C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), CF₃, N₃, (C₁₋₄)alkyl,adamantyl, --SO₂ NH₂, --NHCOCH₃, --NHSO₂ CH₃ or CH₃ SO₂ O-- or by thebridge --O--CH₂ --O--); pyridyl or N-benzylpiperidyl;

or R³ and R⁴ together with the nitrogen atom to which they are boundrepresent pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl orpiperazinyl, whilst the piperazinyl ring may optionally be N-substitutedby methyl, unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl,cyano-substituted phenyl, pyrimidinyl, phenyl(C₁₋₄)alkyl,(C₁₋₄)alkylphenyl or ##STR5## or the salts thereof with physiologicallyacceptable acids or complex-forming agents.

Compounds of formula I form tautomers of formula II ##STR6##

The tautomers can be separated by known methods, e.g. by columnchromatography or selective reduction (NaBH₄ or catalytic reduction).

The compounds of formula II may occur in cis- and/or trans-form:##STR7##

If the structure of a compound is not expressly stated, the mention offormula I should be taken as including structure II as well.

In the definitions used in the text the radicals and groups may beidentical or different, i.e. if one of the above-mentioned substituentsoccurs several times in a particular molecule, the meaning can beselected freely within the scope of the definitions provided.

The term alkyl means C₁₋₆ -alkyl and C₁₋₄ -alkyl radicals which may besubstituted or, as alkyl radicals, are part of a functional group suchas alkoxy or alkylthio. The alkyl radicals include methyl, ethyl,n-propyl, isopropyl, n-butyl, sec.-butyl, isobutyl and tert.-butylradicals as well as the various isomeric pentyl and hexyl radicals, suchas e.g. isopentyl, neopentyl, n-pentyl and n-hexyl radicals.

The above definition thus also applies even when the alkyl radicalitself is substituted and/or is itself part of an alkoxyalkyl-,alkoxycarbonyl-, alkoxy-, alkylthio-, alkylsulphonyl-, monoalkylamino-,alkylmethyl-, alkylthiomethyl- or dialkylamino- group or the alkylradical, as a substituent, is bound to an aromatic heterocyclic orcarbocyclic system.

The halogens are fluorine, chlorine, bromine and iodine, preferablyfluorine, chlorine and bromine and, to a lesser extent, iodine.

C₃₋₆ -cycloalkyl indicates cyclopropane, cyclobutane, cyclopentane andcyclohexane.

C₅₋₆ -cycloalkenes denote e.g. cyclopentene, cyclohexene andcyclohexadiene.

C₃₋₆ -alkynes are the isomeric hexynes, pentynes, butynes and propynes,preferably propargyl.

The C₃₋₆ -alkenes are the isomeric hexenes, pentenes, butenes andpropenes, preferably allyl.

A preferred aspect of the invention consists of compounds of generalformula I wherein A denotes a benzo- or thieno group; wherein, if A isbenzo, m is 2, the R² s are in positions 6 and 7 and independently ofone another represent hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl,Br, I), (C₁₋₄)alkyl, methanesulphonyloxy or methanesulphonamido, or twoadjacent substituents R² may together represent --O--CH₂ --O-- or--O--CH₂ --CH₂ --O--; and if A is thieno, m is zero;

R¹ denotes thienyl or the group ##STR8## wherein R⁷, R⁸ and R⁹independently of one another may represent methyl, ethyl, propyl, phenylor benzyl, whilst not more than 2 of the substituents can simultaneouslyrepresent phenyl or benzyl;

R³ and R⁴ independently of each other represent

(a) hydrogen,

(b) branched or unbranched C₃₋₆ -alkenyl,

(c) branched or unbranched C₃₋₆ -alkynyl, or

(d) branched or unbranched C₁₋₁₂ -alkyl, wherein the alkyl may besubstituted by

hydroxy,

(C₁₋₄)alkoxy,

di(C₁₋₄)alkylamino,

furyl,

pyridyl,

pyrrolidinyl, N-methylpyrrolidinyl,

morpholino,

indolyl,

nitrilo,

thienyl,

adamantyl,

cyclohexyl,

phenoxy,

naphthyloxy or phenyl, whilst this phenyl or the phenyl contained in thephenoxy group may be mono-, di- or trisubstituted by hydroxy,(C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), CF₃, N₃, (C₁₋₄)alkyl,adamantyl, --SO₂ NH₂ or --NHCOCH₃ or by the bridge --O--CH₂ --O--;

or R³ denotes hydrogen and R⁴ denotes cyclohexyl, phenyl, fluorophenyl,pyridyl or N-benzylpiperidyl;

or R³ and R⁴ together with the nitrogen atom to which they are boundrepresent pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl orpiperazinyl, whilst the piperazinyl ring may optionally be N-substitutedby methyl, unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl,pyrimidinyl, phenyl(C₁₋₄)alkyl or ##STR9##

Preferably, A represents the annelated ring systems ##STR10## wherein R²is as hereinbefore defined.

Also preferred according to the invention are compounds I wherein A isindolo and the other substituents are as hereinbefore defined,preferably NR³ R⁴ is either morpholinyl or in NR³ R⁴ R³ is hydrogen andR⁴ is C₁₋₄ -alkyl, which may be substituted as hereinbefore defined.

Of the compounds I wherein A is benzo, the preferred compounds are thosewherein m is 2 and the two R² s independently of each other representmethoxy, hydroxy, benzyloxy, methyl or chlorine or together represent--OCH₂ O--, whilst the two R² s are in positions 6 and 7, particularlythose compounds wherein R² is methoxy, hydroxy, benzyloxy or methyl, andespecially those wherein both R² s are the same and represent hydroxy ormethoxy.

Of the compounds I, the preferred compounds are those wherein R¹ istert.butyl.

Other preferred compounds of formula I are those wherein NR³ R⁴ has oneof the following meanings:

a) in NR³ R⁴, R³ is hydrogen and R⁴ is C₁₋₆ -alkyl;

b) in NR³ R⁴, R³ is hydrogen and R⁴ is branched or unbranched alkynylhaving 3 to 6 (preferably 3) carbon atoms

c) in NR³ R⁴, R³ is hydrogen and R⁴ is branched or unbranched alkylhaving 1 to 4 (preferably 1 to 3, especially 2) carbon atoms, the alkylbeing substituted by

methoxy,

dimethylamino,

pyrrolidinyl, N-methypyrrolidinyl,

morpholino,

thienyl,

adamantyl,

pyridyl,

N-benzylpiperidyl,

cyclohexyl,

phenoxy,

naphthyloxy or 1 or 2 phenyl, whilst this phenyl (if only one phenylgroup is present) or the phenyl contained in the phenoxy group may bemono-, di- or trisubstituted by methoxy, ethoxy, benzyloxy, halogen(particularly Cl, I), CF₃, N₃, methyl, tert.butyl, --SO₂ NH₂, or by thebridge --O--CH₂ --O--;

or R³ denotes hydrogen and R⁴ denotes cyclohexyl, phenyl, fluorophenyl,pyridyl or N-benzylpiperidyl;

d) in NR³ R⁴, R³ and R⁴ independently of each other represent methyl,ethyl, (CH₂)₁₋₄ -phenyl (wherein the phenyl group may be substitutedlike the phenyl group specified in (c)

preferably ##STR11## e) R³ and R⁴ together with the nitrogen atom towhich they are bound denote piperidinyl, morpholinyl, thiomorpholinyl orpiperazinyl, whilst the piperazinyl ring may optionally be N-substitutedby methyl or benzyl;

particularly those wherein NR³ R⁴ has one of the following meanings:

a) in NR³ R⁴, R³ is hydrogen and R⁴ is C₂₋₆ -alkyl;

b) in NR³ R⁴, R³ is hydrogen and R⁴ is CH₂ CCH;

c) in NR³ R⁴, R³ is hydrogen and R⁴ is branched or unbranched C₂₋₄-alkyl, wherein the alkyl is substituted by

methoxy,

dimethylamino,

N-methypyrrolidinyl,

thienyl,

adamantyl,

phenoxy,

naphthyloxy or 1 or 2 phenyl, whilst this phenyl (if there is only onephenyl group present) or the phenyl contained in the phenoxy group maybe mono-, di- or trisubstituted by methoxy, ethoxy, N₃, methyl,tert.butyl or --SO₂ NH₂ ;

d) in NR³ R⁴, R³ and R⁴ independently of each other represent methyl,ethyl, (CH₂)₁₋₄ -phenyl (wherein the phenyl group may be substituted byF) or particularly ##STR12## e) R³ and R⁴ together with the nitrogenatom to which they are bound are piperazinyl, N-substituted by methyl orbenzyl.

Special mention should be made of compounds I wherein NR³ R⁴ has one ofthe following meanings:

a) in NR³ R⁴, R³ is hydrogen and R⁴ is ethyl, tert.butyl or (CH₂)₁ or 2--C(CH₃)₃ ;

b) NR³ R⁴ is NHCH₂ CCH;

c) in NR³ R⁴, R³ is hydrogen and R⁴ is ethyl, propyl or methylpropylwhich is substituted by phenyl, which may be mono-, di- ortrisubstituted by methyl or methoxy or monosubstituted by tert.butyl;

d) in NR³ R⁴, R³ and R⁴ are identical, namely ##STR13## e) NR³ R⁴ is##STR14## particularly those wherein R³ is hydrogen or(C₁₋₄)alkyl-phenyl and R⁴ is (C₁₋₄)alkyl-phenyl, whilst in these groupsC₁ -alkyl is preferably present and phenyl is mono-substituted byhalogen (preferably Cl or F), CF₃, methoxy or ethoxy, this substituentpreferably being in the o-position.

The compounds of formula I may be prepared by methods known per se,preferably analogously to the method described in German PatentApplication P 37 18 570.5, EP 358 957, EP 37 934, EP 251 794 and EP 288048.

In the presence of a condensing agent, a malonic acid diamide of generalformula IV ##STR15## wherein R¹, R², R³, R⁴ and m are as hereinbeforedefined and Ar represents phenyl, indolyl or 2- or 3-thienyl, may becyclised to obtain the corresponding compounds.

Suitable condensing agents for this process are strong Lewis acids suchas phosphorusoxychloride, phosphoruspentachloride,phosphorustrichloride, phosphoruspentoxide, titanium tetrachloride,boron trifluoride, tin tetrachloride, as well as inorganic acids such aspolyphosphoric acid, sulphuric acid, fluorosulphonic acid andhydrofluoric acid, or mixtures of condensing agents such as a mixture ofphosphorusoxychloride and phosphoruspentachloride, or a mixture ofphosphoruspentoxide and (C₁₋₄)alkylsulphonic acid, e.g. with a P₂ O₅ --content of about 10% by weight.

The cyclisation may be carried out in the presence or absence of asolvent. Any inert solvents are suitable provided that they havesufficient solubility for the reactants and a high enough boiling point,e.g. benzene, alkylbenzenes (e.g. toluene, xylene), chlorobenzenes,chloroform, acetonitrile and decaline. According to a preferredembodiment of the process the condensing agent used isphosphorusoxychloride in admixture with acetonitrile or a mixture of(C₁₋₄)alkylsulphonic acid and phosphoruspentoxide, without the additionof solvents.

Preferably, the cyclisation is carried out withphosphorusoxychloride/acetonitrile or in difficult cases with a mixtureof phosphoruspentoxide and C₁₋₄ -alkylsulphonic acid (preferablymethanesulphonic acid). The reaction can be carried out in a widetemperature range, preferably with heating to 50° C. up to the boilingpoint of the reaction mixture.

The necessary reaction period will be between 2 and 15 hours dependingon the starting compound IV.

The 3-thiophenmalonic acid required for this preparation is commerciallyavailable. The 2-thiophenmalonic acid may be prepared by methods knownper se (e.g. from 2-thiophenacetic acid using the carbonate method orfrom 2-thiophenbromide and diethylmalonate).

The compounds of formula I are bases and can be converted in the usualway with inorganic or organic acids and salts and complex-forming agentsinto any desired physiologically acceptable adducts (salts).

Acids suitable for salt formation include for example hydrochloric,hydrobromic, hydriodic, hydrofluoric, sulphuric, phosphoric, nitric,acetic, propionic, butyric, caproic, valeric, oxalic, malonic, succinic,maleic, fumaric, lactic, tartaric, citric, malic, benzoic,p-hydroxybenzoic, phthalic, cinnamic, salicylic, ascorbic,methanesulphonic acid and the like.

The compounds may be administered by oral, parenteral or topical route.The desired therapeutic dose depends on the indication and formulationused and can be determined experimentally. Suitable forms include, forexample, tablets, capsules, suppositories, solutions, syrups, emulsions,aerosols or dispersible powders. Tablets may be produced, for example,by mixing the active substance or substances with known excipients, e.g.inert diluents such as calcium carbonate, calcium phosphate or lactose,disintegrants such as corn starch or alginic acid, binders such asstarch or gelatine, lubricants such as magnesium stearate or talc and/oragents for obtaining delayed release, such as carboxypolymethylene,carboxymethylcellulose, cellulose acetate phthalate or polyvinylacetate.The tablets may also consist of several layers.

Coated tablets may be produced analogously by coating cores made in thesame way as the tablets with substances conventionally used for tabletcoatings, e.g. collidone or shellac, gum arabic, talc, titanium dioxideor sugar. In order to obtain delayed release or avoid incompatibilities,the core may also consist of several layers. Similarly, the tabletcoating may consist of several layers to achieve delayed release, whilstthe excipients mentioned for the tablets may be used.

Syrups containing the active substances or combinations of activesubstances according to the invention may additionally contain asweetener such as saccharin, cyclamate, glycerol or sugar as well as aflavour enhancer, e.g. a flavouring such as vanillin or orange extract.They may also contain suspension adjuvants or thickeners such as sodiumcarboxymethylcellulose, wetting agents, e.g. condensation products offatty alcohols with ethylene oxide or preservatives such asp-hydroxybenzoates.

Injectable solutions are produced in the usual way, e.g. by addingpreservatives such as p-hydroxybenzoates or stabilisers such as alkalimetal salts of ethylene diamine tetraacetic acid, and are thentransferred into injection vials or ampoules.

Capsules containing one or more active substances or combinations ofactive substances may be prepared for example by mixing the activesubstances with inert carriers such as lactose or sorbitol andencapsulating them in gelatine capsules.

Suitable suppositories may be produced for example by mixing withcarriers provided for this purpose, such as neutral fats orpolyethyleneglycol or derivatives thereof.

The compounds may be administered both enterally and parenterally. Aproposed dose for oral use is 0.1 to 500 mg of active substance per doseand from 0.05 to 150 mg per dose for intravenous administration. Thedesired therapeutic dose depends on the indication and formulation usedand can be determined experimentally.

The pharmaceutical compositions are suitable for oral or parenteral andpossibly topical application. The chief formulations used are plain orcoated tablets, ampoules and syrups. The single dose using theseformulations is between 1.0 and 200 mg, preferably 20 to 50 mg per 75 kgof body weight. Generally, 1 to 3 single doses are required per day,depending on the gravity of the case.

The following Examples serve to illustrate the invention:

EXAMPLE 1

1. Monoethyl tert.butyl malonate

At ambient temperature over a period of 30 minutes, a solution of 7.6 gof KOH (85% strength) in 50 ml of water is added dropwise, withstirring, to a mixture of 21.6 g of diethyltert.butylmalonate, 50 ml ofethanol and 50 ml of water. After 15 hours the ethanol is distilled offin vacuo. 300 ml of CH₂ Cl₂ are added to the residue after cooling. Thisis acidified with a solution of 13.6 g of KHSO₄ in 100 ml of H₂ O,whilst cooling with ice, and the aqueous phase is extracted severaltimes with CH₂ Cl₂. The combined organic phases are washed with water,dried over Na₂ SO₄ and concentrated by evaporation in vacuo at a bathtemperature of 30° C. 15.35 g (=81.7% of theory) of monoethylesterremain (bright yellow oil).

2. Monoethyl tert.butyl malonate-N- 2-(3,4-dimethoxyphenyl)ethyl!-amide

To a solution of 15.35 g of monoethyl tert.butyl malonate in 200 ml ofanhydrous CH₂ Cl₂ are stirred, at ambient temperature, 15.88 g ofN,N'-carbonyldiimidazole in small batches. After 30 minutes, 14.8 g of2-(3,4-dimethoxyphenyl)ethylamine are added. After a further 15 hoursthe solvent is distilled off in vacuo. The residue is mixed with 200 mlof water, acidified with 2N HCl and extracted with ethyl acetate. Theorganic phase is washed with water and concentrated by evaporation afterdrying over Na₂ SO₄. The residue is triturated with petroleum ether (40°to 60° C.) and brought to crystallisation.

Yield: 24.7 g (86.2% of theory); m.p. 68°-70° C.

3. tert.Butylmalonic acid-mono-N- 2-(3,4-dimethoxyphenyl)-ethyl!amide

42 g of esteramide (see above) and 180 ml of 1N NaOH are refluxed for 2hours. After cooling and filtering, the solution is extracted with etherand acidified by the addition of 50 ml of 4N HCl. As the solution isleft to stand, crystals are precipitated out. They are suction filteredand dried at 40° to 50° C.

Yield 35.4 (91.3% of theory). M.p. 103°-104° C.

4. tert.Butylmalonic acid-N-2-(3,4-dimethoxyphenyl)-ethyl!-N'-(3,3-diphenylpropyl)-diamide

At ambient temperature, 2.1 g of N,N'-carbonyldiimidazole are added to asolution of 3.23 g of tert.butylmalonic acid monoamide (from Example 3)in 50 ml of anhydrous CH₂ Cl₂. After 30 minutes, 2.11 g of3,3-diphenylpropylamine are added. After 15 hours' standing at ambienttemperature the solvent is distilled off. The residue is mixed withwater, acidified with 2N HCl and extracted with ethyl acetate. Theorganic phase is washed with water, dried over Na₂ SO₄ and evaporateddown in vacuo. The residue (4.9 g ≅95% of theory) is used in thecyclisation reaction without any further purification.

5.(R,S)-(3,4-Dihydro-6,7-dimethoxyisoquinolin-1-yl)-2-tert.butyl-N-(3,3-diphenylpropyl)-acetamide

A mixture of 5.15 g of tert.butylmalonic acid diamide (from Example 4),1.9 ml of POCl₃ and 35 ml of acetonitrile is refluxed for 2 hours. Aftercooling, it is poured on to ice water, made alkaline with saturated sodasolution and extracted with ethyl acetate. The organic phase is washedwith water, dried over Na₂ SO₄ and evaporated down. The residue isdissolved in acetone, converted into the hydrochloride with thecalculated quantity of ethereal hydrochloric acid and crystallised bytrituration.

Yield: 4.45 g (83.6% of theory); m.p. 147°-148° C.

EXAMPLE 2 3-Thienylmalonic acid-N- 2-(3,4-dimethoxyphenyl)ethyl!-amide##STR16##

15.1 g (40 Mmol) of monoethyl-3-thienylmalonate-N-2-(3,4-dimethoxyphenyl)ethyl!-amide are dissolved in 150 ml of methanoland 100 ml of dioxane and added dropwise to 42 ml (42 Mmol) of 1N NaOH,with stirring and cooling with ice. The mixture is stirred for a further2 hours at ambient temperature, the organic solvents are distilled offin vacuo and the residue is distributed between water and CH₂ Cl₂.

The aqueous phase is acidified with 10% citric acid, with cooling andstirring, and extracted with CH₂ Cl₂. After washing with water,saturated NaCl solution and drying over MgSO₄, the solvent is distilledoff in vacuo and a residue of 12.7 g is obtained. 11.7 g (83.7% oftheory) of the title compound are obtained by recrystallisation frommethylene chloride/methanol/ether.

3-Thienylmalonic acid-N-2-(3,4-dimethoxyphenyl)ethyl!-N'-dibenzyl-diamide ##STR17##

To a solution of 3.75 g (11 Mmol) of 3-thienylmalonic acid-N-2-(3,4-dimethoxyphenyl)-ethyl!-amide in 100 ml of absolutetetrahydrofuran are added, at 5° C., with stirring, 1.78 (11 Mmol) ofcarbonyldiimidazole, in small batches. The reaction mixture is stirredfor 30 minutes at ambient temperature and then 2.17 g (11 Mmol) ofdibenzylamine are added. After 16 hours stirring at ambient temperaturethe mixture is evaporated down and the residue is distributed betweenethyl acetate and water. The organic phase is washed successively withwater, 5% KHSO₄ solution, saturated NaHCO₃ solution, water and saturatedNaCl solution, dried over MgSO₄ and the mixture of solvents is distilledoff in vacuo. The residue is recrystallised from a little ether.

Yield: 5.1 g (87.7% of theory) of the title compound are obtained.

(R,S)-(3,4-Dihydro-6,7-dimethoxyisoquinolin-1-yl)-2-(3-thienyl)-N,N-dibenzyl-acetamide##STR18##

5.1 g (96 Mmol) of 3-thienylmalonic acid-N-2-(3,4-dimethoxyphenyl)ethyl!-N'-dibenzyl-diamide are combined with 4.41g (28.8 Mmol) of phosphorusoxychloride in 50 ml of acetonitrile(analytical grade) and the mixture is refluxed for 1 hour under an N₂atmosphere. After cooling, 150 ml of ethyl acetate are added, themixture is neutralised with saturated NaHCO₃ solution, washed with waterand saturated NaCl solution, dried over MgSO₄ and the solvents aredistilled off in vacuo. The residue is dissolved in 10 ml of absoluteacetone, 900 mg (10 Mmol) of oxalic acid are added and the salt isprecipitated in crystalline form after the addition of about 50 ml ofabsolute diethylether.

Yield: 4.8 g (83.3% of theory) of the title compound in the form of theoxalate; m.p.: 128°-130° C.

The following Table lists examples of compounds according to theinvention. These compounds may be prepared analogously to the methodsdescribed above.

                                      TABLE 1                                     __________________________________________________________________________     ##STR19##               M.p. (°C.)                                                                  Saltform                                                                           Tautomer Structure                         __________________________________________________________________________    X =                                                                               ##STR20##            76   Fu(1.5)                                             ##STR21##            105  Fu                                                  ##STR22##            152  Fu(1.5)                                             ##STR23##            230  Cl                                                  ##STR24##            138  Fu(1.5)                                             ##STR25##            112  Fu                                                  ##STR26##                                                                     ##STR27##                                                                     ##STR28##            98   Fu(1.5)                                             ##STR29##                                                                     ##STR30##            209  Cl                                                  ##STR31##                                                                     ##STR32##            215  Cl                                                  ##STR33##                                                                     ##STR34##                                                                     ##STR35##                                                                     ##STR36##                                                                     ##STR37##            174  Fu                                                  ##STR38##            208  Fu                                                  ##STR39##            232  Cl                                                  ##STR40##            165  Cl                                                  ##STR41##            133  Fu(1.5)                                                                            I                                             NH(CH.sub.2).sub.9CH.sub.3                                                                          147  Cl                                                  ##STR42##            210  Cl                                                  ##STR43##            223  Cl                                                  ##STR44##                                                                     ##STR45##                                                                     ##STR46##                                                                     ##STR47##            209  Cl   I                                              ##STR48##            187  Fu(1.5)                                             ##STR49##                                                                     ##STR50##                                                                     ##STR51##            102  Fu(1.5)                                             ##STR52##            201  Fu                                                  ##STR53##            212  Cl                                                  ##STR54##            145  Fu(1.5)                                             ##STR55##            145  Fu(1.5)                                             ##STR56##            192  Fu(1.5)                                             ##STR57##            158  Fu(2)                                               ##STR58##            209  Cl                                                  ##STR59##            170  Cl                                                  ##STR60##            138  Cl                                                  ##STR61##            188  Cl                                                  ##STR62##            222  Cl                                                  ##STR63##            144  Fu(1.5)                                             ##STR64##            180  Fu(1.5)                                             ##STR65##            210  Fu(1.5)                                             ##STR66##            222  Cl                                              __________________________________________________________________________

                                      TABLE 2                                     __________________________________________________________________________     ##STR67##               Saltform                                                                           M.p. (°C.)                               __________________________________________________________________________    X =                                                                               ##STR68##            OX   125-135 (decomp.)                                   ##STR69##            BS   161-163                                             ##STR70##                                                                     ##STR71##                                                                     ##STR72##            BS   118-120                                             ##STR73##                                                                     ##STR74##                                                                     ##STR75##                                                                     ##STR76##                                                                     ##STR77##                                                                     ##STR78##            BS   128-130                                             ##STR79##                                                                     ##STR80##            BS   185-187                                             ##STR81##                                                                     ##STR82##            OX   128-130                                             ##STR83##                                                                     ##STR84##                                                                     ##STR85##                                                                     ##STR86##            OX   70-80 (decomp.)                                     ##STR87##            BS   165-167                                             ##STR88##            BS   102-104                                             ##STR89##            OX   124-127                                            NH(CH.sub.2).sub.9CH.sub.3                                                                          OX   121-122                                             ##STR90##                                                                     ##STR91##                                                                     ##STR92##                                                                     ##STR93##                                                                     ##STR94##            OX   107-112                                             ##STR95##                                                                    NH.sub.2              OX   121-140 (decomp.)                                   ##STR96##            OX   amorph                                              ##STR97##            OX   80-100 (decomp.)                                    ##STR98##                                                                 __________________________________________________________________________

                  TABLE 3                                                         ______________________________________                                         ##STR99##                                                                    X:                   Saltform M.p. (°C.)                               ______________________________________                                         ##STR100##          BS       amorph                                          ______________________________________                                         OX: Oxalate                                                                   BS: free base                                                                 Decomp.: Decomposition                                                   

The present invention also relates to the use of these new compounds.

The compounds are valuable in the treatment of degenerative and necroticdiseases of the brain. It is also possible to provide preventativetreatment for patients who are at risk from such diseases. The effect ofthe compounds is not based on an improvement in the blood flow throughthe tissues. The compounds are therefore suitable for a new kind oftreatment of epilepsy and Alzheimer's disease and particularly fortreating patients who have suffered a stroke or are at risk of sufferinga stroke.

The present invention further relates to the use of the above compoundsfor preparing agents for the treatment of chronic inflammatoryprocesses, ulcerative colitis and Crohn's disease and agents with anantiproliferative activity. The effect of the compounds can be explainedby their inhibition of the unselective cation channels (UCC).

The pathophysiology of chronic bronchial asthma is based on inflammatoryprocesses which are mediated by the activation of inflammatory cells.(BARNES, 1987; SEIFERT and SCHULTZ, 1991).

The receptor-regulated activation of inflammatory cells (e.g.neutrophilic granulocytes and mast cells or the permanent cell linesHL-60 cells or sensitised RBL cells, i.e. those charged withgammaglobulin E) is inhibited, irrespective of the nature of thestimulating agonists (e.g. endothelin, PAF, leukotrienes, chemotacticalpeptide fMLP or antigen against sensitised mast cells) by blockers ofunselective cation channels (UCC) (RINK, 1990). Through these channelsextracellular calcium, which is responsible for the persistence ofreceptor-mediated cell activations, enters the cells (PUTNEY, 1990). Ifthis supply of calcium is interrupted this results in a blockade of theactivation of inflammatory cells.

Conventional calcium antagonists of the dihydropyridine orphenylalkylamine type do not inhibit either UCCs or inflammatoryprocesses (WELLS et al., 1986).

As a measurement of the cell activation or as a measurement of theinhibition thereof by UCC blockers, the kinetics of the cytoplasmiccalcium ion concentration in fura-2-charged cells is quantifiedfluorometrically using the method described by GRYNKIEWICZ et al.(1985). This procedure has proved a reliable screening method, withinthe scope of the invention, for detecting UCC blockers.

So-called functional THAPSIGARGIN inhibition has proved suitable for thespecific characterisation of blockers of the unselective cationchannels. THAPSIGARGIN is a tumour promoter described by THASTRUP et al.(Proc. Natl. Acad. Sci. (U.S.A.), 87, 2466-2470, 1990) which selectivelyand irreversibly inhibits the Ca²⁺ -ATPase of intracellular IP₃-sensitive Ca²⁺ -stores. Consequently the Ca²⁺ -stores are rapidlydepleted. As described by J. PUTNEY (Calcium, 11, 611-624, 1990) thedepletion of these stores constitutes the physiological stimulation foropening up unselective cation channels in the cell membrane. The resultof this is a massive influx of Na⁺ and Ca²⁺ into the cell. Because ofthese properties, Thapsigargin is suitable as an indirect stimulator foragonist- and IP₃ -independent opening up of the unselective cationchannels.

Within the scope of the present invention the Thapsigargin stimulationof unselective cation channels has been carried out successfully on HL60 cells (human leukaemia cells), on hippocampal and cortical neuronecells and on RBL-cells (rat basophilic lymphoma cells) and in this waythe existence of these channels in particular cell lines wasdemonstrated.

The cytoplasmic Ca²⁺ concentration ( Ca²⁺ !_(i)) plays an important partin the cell proliferation and in tumour growth (for a summary see L. R.ZACHARSKI, Journal of Medicine 19: 145-177, 1988). In particular, theCa²⁺ -influx into the cell stimulated by receptor activation withconsecutive inositoltriphosphate-(IP₃ -)-mediation would appear to be ofcrucial importance for oncogenic cell proliferation (U. KIKKAWA and Y.NISHIZUKA, Ann. REV. CELL. BIOL. 2: 149-178, 1986). This mechanism alsoplays a part in the formation of metastases and in "Multi-DrugResistance". (For a summary see the above-mentioned publication by L. R.ZACHARSKI, J. MED. 19: 145-177, 1980).

This hypothesis is supported by the fact that Thapsigargin, as anindirect stimulator of the unselective cation channels (UCC) not onlyleads to a Ca²⁺ -overload in the cell but is also a highly effectivetumour promoter. (V. THASTRUP et al. Proceedings of the NATL. Acad. Sci:(U.S.A.) 87: 2466-2470, 1990). The blockade of the Ca²⁺ -influx by theUCC leads to normalisation of the intracellular Ca-ion concentration andhence to inhibition of tumour growth etc.

Conventional calcium antagonists do not inhibit these UCC. It has beenfound, surprisingly, that the compounds according to this inventioninhibit the influx of calcium into the cell through the UCC.

As shown by S. H. MURCH et al. (Lancet 339 : 381-385, 15. Feb. 1992)endothelin I plays an important pathophysiological role in inflammatoryintestinal diseases such as ulcerative colitis and Crohn's disease.Using immunohistochemical methods it has been shown that patients withCrohn's disease in the region of the submucosa and patients withulcerative colitis in the region of the lamina propria of the epitheliumof the large intestine show significantly and greatly increasedconcentrations of endothelin I compared with healthy normal people. Itis assumed that the local secretion of endothelin causes massivevasospasms with consecutive disseminated ischaemia with microinfarctswhich are regarded as the actual cause of the above diseases. Thevasospasmogenic effectiveness of endothelin is explained by a Ca²⁺-overload of vascular myocytes. Endothelin primarily triggers an IP₃-mediated intracellular release of Ca²⁺ which is followed by a massivetransmembranal Ca² +-entry through dihydropyridine-insensitive channels.(M. S. Simonson et al. Clin. Invest. Med. 14: 499-507, 1991; T. Masakai,J. Cardiovasc. Pharmacol. 13:Suppl. 5, S1-S4, 1989; D. W. Hay, R. J.Pharmacol. 100: 383-392, 1990). These channels are unselective cationchannels which have also been briefly described as existing in cells ofthe large intestine mucosa. (Chr. Siemer and H. Gogelein, Europ. J.Physiol. 420: 319-328, 1992).

The endothelin-stimulated activation of fura-2-charged human leukaemiacells (HL 60 cells) has proved a suitable screening model for detectingfunctional endothelin antagonists. In conformity with G. GRYNKIEWICZ etal. (J. Biol. Chem. 260:3440-3450, 1985) the intracellular Ca²⁺-concentration in the cytoplasm of HL 60 cells (suspensions) can bemonitored by spectrofluorometry and quantified as a measurement of cellactivation by endothelin. The stimulation was effected by adding 0.1 mMendothelin and could be inhibited in a dosage-dependent manner by meansof the substances according to the invention.

The functional endothelin antagonism of the substances according to theinvention is mediated through a blockade of the unselective cationchannels.

Consequently, detection of a functional Thapsigargin-antagonism onRBL-hm1 cells is also a suitable screening method for functionalendothelin antagonists.

Carrying out the investigation:

For screening purposes, fura-2-charged adhesive RBL-hm 1 cells arestimulated with 0.1 μM Thapsigargin in a Ca²⁺ -free incubation medium.After 4 minutes, extracellular Ca²⁺ is restored to a concentration of1.5 mM and, using the fura-2-fluorescence, the excessive increase in thecytoplasmic Ca²⁺ -concentration caused by a massive transmembranal Ca²⁺-entry through unselective cation channels is recorded.

This entry is to be inhibited solely by unselective cation channelblockers in a dosage-dependent manner. Neither conventional calciumantagonists nor specific blockers of agonists which stimulate the IP₃-turnover are able to inhibit the transmembranal Ca²⁺ -entry triggeredindirectly by Thapsigargin. The compounds of the present invention aredistinguished by their inhibition of UCC.

The fluorometric calcium measurement in the cytoplasm of individualadhering RBL-hm1 cells is carried out analogously to the methoddescribed by KUDO and OGURA (1986) for neuronal cells. An AXIOVERT 35fluorescence microscope made by ZEISS is used in conjunction with animaging system made by HAMAMATSU, consisting of the ICMS-imageprocessing system, residual light camera with control unit and imageintensifier DVS 3000.

The kinetics of the cytoplasmic Ca²⁺ -concentration is recordedcontinuously as a concentration/time curve after the cell activationstimulated by Thapsigargin (0.1 μM). The curves of two activated cellcultures are compared in the presence and absence of 10 μM testsubstance. The area under these curves (area under the curve=AUC) isintegrated and recorded as a measurement of cell activation. Theinhibitory potency of the UCC-blockers tested is determined using thefollowing equation: ##EQU1## % H=the percentage inhibition of thecalcium entry through unselective cation channels which is stimulatedand inhibited by 10 μM of test substance.

AUC_(inh) =area under the curve recorded in the presence of thestimulant plus 10 μM inhibitory test substance.

AUC control=area under the curve which is recorded only after theaddition of the stimulant.

Literature relating to the above explanations:

BARNES P. J., I. W. RODGER and N. C. THOMSON Pathogenesis of asthma, in"ASTHMA, basic mechanisms and clinical management" ED by P. J. BARNES;ACADEMIC PRESS, LONDON, 1988

GRYNKIEWICZ G., M. POENIE and R. Y. TSIEN A new generation of Ca²⁺-indicators with greatly improved fluorescence properties J. BIOL. CHEM.260: 3440-3450, 1985

HIDE, M. and M. A. BEAVEN Calcium influx in a rat mast cell (RBL-2H3)line J. BIOL. CHEM. 266 15221-15229, 1991

KUDO, Y. and A. OGURA Glutamate-induced increase in intracellular Ca²⁺-concentration in isolated hippocampal neurones

BR. J. PHARMACOL. 89: 191-198; 1986

PUTNEY, J. W., jr. Capacitative Calcium entry revised CELL CALCIUM 11:611-624, 1990

RINK, T. J. Receptor-mediated calcium entry FEBS LETT. 268: 381-385,1990

SEIFERT, R. and G. SCHULTZ The superoxide forming NADPH oxidase ofphagocytes: An enzyme system regulated by multiple mechanism REV.PHYSIOL. BIOCHEM. PHARMACOL., Vol. 117, SPRINGER VERL., 1991

WELLS, E., C. G. JACKSON, S. T. HARPER, J. MANN and R. P. EAOYCharacterization of primate bronchoalveolar mast cells II, inhibition ofhistamine, LTC₄ and PGF_(2a) release from primate bronchoalveolar mastcells and a comparison with rat peritoneal mast cells J. IMMUNOL. 137:3941-3945, 1986.

Results of measurement

The percentage inhibition of UCC after Thapsigargin stimulation (0.1 μMThapsigargin) in RBL-hm 1 cells is given. The uniform concentration ofthe test substances is 10⁻⁵ mol or 10⁻⁶ mol)

                                      TABLE 4                                     __________________________________________________________________________     ##STR101##              Saltform                                                                           % H (10.sup.-5 M)                                                                    % H (10.sup.-6 M)                        __________________________________________________________________________    X =                                                                               ##STR102##           Fu(1.5)     71.9                                         ##STR103##           Fu          57.3                                         ##STR104##           Fu(1.5)     71.2                                         ##STR105##           Cl          66.4                                         ##STR106##           Fu(1.5)     53.3                                         ##STR107##           Fu          70.7                                         ##STR108##                                                                    ##STR109##                                                                    ##STR110##           Fu(1.5)                                                                            60.5                                                ##STR111##                                                                    ##STR112##           Cl          64.6                                         ##STR113##                                                                    ##STR114##           Cl          70.1                                         ##STR115##                                                                    ##STR116##                                                                    ##STR117##                                                                    ##STR118##                                                                    ##STR119##           Fu          77.2                                         ##STR120##           Fu          75.7                                         ##STR121##           Cl          24.8                                         ##STR122##           Cl          64.4                                         ##STR123##           Fu(1.5)     74.4                                        NH(CH.sub.2).sub.9CH.sub.3                                                                          Cl          69.7                                         ##STR124##           Cl          45.8                                         ##STR125##           Cl          80.8                                         ##STR126##                                                                    ##STR127##                                                                    ##STR128##                                                                    ##STR129##           Cl          64.9                                         ##STR130##           Fu(1.5)                                                                            66.8                                                ##STR131##                                                                    ##STR132##                                                                    ##STR133##           Fu(1.5)     73.8                                         ##STR134##           Fu          29.2                                         ##STR135##           Cl          72.7                                         ##STR136##           Fu(1.5)     28.4                                         ##STR137##           Fu(1.5)     43.2                                         ##STR138##           Fu(1.5)     16.9                                         ##STR139##           Fu(2)       35.7                                         ##STR140##           Cl          53.3                                         ##STR141##           Cl   100.0  60.3                                         ##STR142##           Cl   93.5   47.3                                         ##STR143##           Cl   99.0   63.7                                         ##STR144##           Cl   96.0   65.4                                         ##STR145##           Fu(1.5)                                                                            91.6   65.7                                         ##STR146##           Fu(1.5)                                                                            65.8                                                ##STR147##           Fu(1.5)     51.1                                         ##STR148##           Cl          61.0                                     __________________________________________________________________________

                                      TABLE 5                                     __________________________________________________________________________     ##STR149##              Saltform                                                                           % H (10.sup.-5 M)                               __________________________________________________________________________    X =                                                                               ##STR150##           OX   20.85                                               ##STR151##           BS   59.94                                               ##STR152##                                                                    ##STR153##                                                                    ##STR154##           BS   37.83                                               ##STR155##                                                                    ##STR156##                                                                    ##STR157##                                                                    ##STR158##                                                                    ##STR159##                                                                    ##STR160##           BS   36.41                                               ##STR161##                                                                    ##STR162##           BS   39.17                                               ##STR163##                                                                    ##STR164##           OX   90.9                                                ##STR165##                                                                    ##STR166##                                                                    ##STR167##                                                                    ##STR168##           OX   22.62                                               ##STR169##           BS   14.6                                                ##STR170##           BS   25.76                                               ##STR171##           OX   50.61                                              NH(CH.sub.2).sub.9CH.sub.3                                                                          OX   47.09                                               ##STR172##                                                                    ##STR173##                                                                    ##STR174##                                                                    ##STR175##                                                                    ##STR176##           OX   28.14                                               ##STR177##                                                                   NH.sub.2              OX                                                       ##STR178##           OX   6.8                                                 ##STR179##           OX   52.26                                               ##STR180##                                                                __________________________________________________________________________

                  TABLE 6                                                         ______________________________________                                         ##STR181##                                                                                      Saltform                                                                             % H (10.sup.-5 M)                                   ______________________________________                                         ##STR182##          BS       27.01                                           ______________________________________                                    

The functional antiinflammatory effectiveness can be demonstrated bymeans of the following test:

Individual RBL-2H3-cells (a tumour cell line related to the mast cells)adhering to glass slides are used.

The cultivation and attachment of the RBL-2H3-cells are carried out bythe method described by HIDE and BEAVEN (1991). In order to sensitisethe adhesive RBL-2H3-cells the cells are incubated for 2 hours atambient temperature with a 1:2000 diluted commercial gammaglobulinE-solution against a dinitrophenol-bovine serum albumin complex(DNP-BSA-antigen). The cells are then washed. By the addition of 0.1 mlof DNP-BSA-solution (10 μg/ml) there is a massive immunological cellactivation which is mediated by a cytoplasmic Ca²⁺ -overload. Thefluorometric calcium measurement in the cytoplasm of individual adheringRBL-2H3-cells is carried out analogously to the method described by KUDOand OGURA (1986) for neuronal cells, which is also explainedhereinbefore in this specification.

The comparison used in these investigations is (10 μM) chromoglycatewhich brings about an approximately 50% inhibition of theantigen-induced cell activation.

In this test the above-mentioned compounds demonstrate % H values whichare comparable with the values specified hereinbefore.

Tests on microcultures of various human tumour cell lines using thetetrazolium assay in order to determine the antiproliferative effect ofthe substances according to the invention surprisingly showed that thecompound tested was 5 to 100 times more potent than the comparisonsubstance Verapamil.

The antiproliferative effectiveness of the test substances wasdetermined by means of the MTT test described by MOSMANN (J. IMMUNOL.METH. 65: 55-63, 1983), DENIZOT et al. (J. IMMUNOL. METH. 89: 271-277,1986) and J. ELIASON et al. (INT. J. CANCER 46: 113-117, 1990). (MTT=3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide! producedby CHEMICON Inc. El Segundo, Calif., U.S.A.). This indicator ismetabolised only by living cells with intact mitochondria into a blueformazane product. The following human tumour cell lines were used inour test: A 549 (adenocarcinoma of the lung), A 431 (epidermal carcinomaof the vulva), PC 3 (adenocarcinoma of the prostate), SK BR 3(adenocarcinoma of the breast), HT 29 (CX1 1) (adenocarcinoma of thecolon) and K 562 (chronic myeloid leukaemia cell).

The test was carried out on microtitre plates. Each well contained 100μl of a cell suspension (0.2×10⁶ cells per ml). The incubation mediumused was RPMI 1640 with 10% heat-inactivated foetal calves' serum and 50μg/ml of gentamycin. The cell suspensions were incubated for 0, 24, 48or 72 hours in air with a humidity at saturation point in a CO₂ (5%)/air (95%) mixture at 37° C., incubated in the presence and absence ofvariable concentrations of antiproliferative test substances. The testsubstances were dissolved in DMSO (final dilution: 0.1%). Then 10 μl ofMTT-solution (3 mg/ml) were added, followed after 3 hours by 100 μl ofan isopropanol solution containing 0.08N HCl. After a further hour, thelight absorption at 570 nm (comparative wavelength 630 nm) wasdetermined in a microplate reader. The light absorption is directlyproportional to the number of living cells. The half-maximum inhibitoryconcentrations of the substances tested were 1 μg/ml.

The vasospasmolytic effectiveness of the above-mentioned functionalendothelin and Thapsigargin antagonists were confirmed on an isolatedblood vessel preparation: coronary perfusion was continuouslyquantified, on retrogressively perfused, spontaneously beatingLANDENDORFF hearts taken from rats, by means of electromagnetic flowmeasurement (apparatus supplied by Hugo Sachs Elektronik, MARCH). Thismeasuring apparatus could be used to record the extent, duration andpattern of vascular spasms with a high degree of accuracy. If perfusionis carried out with 100 nM endothelin concentration, the coronaryperfusion flow is reduced from 11 to 5 ml/min. The restriction inperfusion can be reversed by means of the substances according to theinvention. The potencies of the compounds according to the inventionwith regard to Thapsigargin inhibition on fura-2-charged RBL-hm1-cellsor the effectiveness of endothelin-inhibition on fura-2-charged HL 60cells correlates clearly with the vasospasmolytic effectiveness of thetest substances detected on the Langendorff preparation. It can beconcluded from this that, underlying the vasospasmoIytic endothelinantagonism of the substances tested, there is a blockade of theunselective cation channels.

Examples of Pharmaceutical Preparations

    ______________________________________                                        a) Coated tablets                                                             ______________________________________                                        1 tablet core contains:                                                       Active substance of general formula I                                                              30.0       mg                                            Lactose              100.0      mg                                            Corn starch          75.0       mg                                            Gelatine             3.0        mg                                            Magnesium stearate   2.0        mg                                                                 210.0      mg                                            ______________________________________                                    

Preparation

The active substance mixed with lactose and corn starch is granulatedwith a 10% aqueous gelatine solution through a 1 mm mesh screen, driedat 40° C. and rubbed through a screen once more. The granules thusobtained are mixed with magnesium stearate and compressed. The coresproduced in this way are coated in the usual manner with a coatingconsisting of an aqueous suspension of sugar, titanium dioxide, talc andgum arabic. The finished coated tablets are polished with beeswax.

    ______________________________________                                        b) Tablets                                                                    ______________________________________                                        Active substance of general formula I                                                              30.0       mg                                            Lactose              100.0      mg                                            Corn starch          70.0       mg                                            Soluble starch       7.0        mg                                            Magnesium stearate   3.0        mg                                                                 210.0      mg                                            ______________________________________                                    

Preparation

The active substance and magnesium stearate are granulated with anaqueous solution of the soluble starch, the granules are dried andintimately mixed with lactose and corn starch. The mixture is thencompressed into tablets weighing 210 mg.

    ______________________________________                                        c) Capsules                                                                   ______________________________________                                        Active substance according to formula I                                                             20.0      mg                                            Lactose               230.0     mg                                            Corn starch           40.0      mg                                            Talc                  10.0      mg                                                                  300.0     mg                                            ______________________________________                                    

Preparation

The active substance, lactose and corn starch are first combined in amixer and then in a grinding machine. The mixture is returned to themixer, thoroughly combined with the talc and mechanically packed intohard gelatine capsules.

    ______________________________________                                        d) Tablets                                                                    ______________________________________                                        Active substance according to the invention                                                          40.0      mg                                           Lactose                100.0     mg                                           Corn starch            50.0      mg                                           Colloidal silica       2.0       mg                                           Magnesium stearate     3.0       mg                                           total                  195.0     mg                                           ______________________________________                                    

Preparation

The active substance is mixed with some of the excipients and granulatedwith a solution of the soluble starch in water. After the granules havedried the remaining excipients are added and the mixture is compressedto form tablets.

    ______________________________________                                        e) Coated tablets                                                             ______________________________________                                        Active substance according to the invention                                                          20.0      mg                                           Lactose                100.0     mg                                           Corn starch            65.0      mg                                           Colloidal silica       2.0       mg                                           Soluble starch         5.0       mg                                           Magnesium stearate     3.0       mg                                           total                  195.0     mg                                           ______________________________________                                    

Preparation

The active substance and excipients are compressed to form tablet coresas described in Example a) and these are then coated in the usual waywith sugar, talc and gum arabic.

    ______________________________________                                        f) Suppositories                                                              ______________________________________                                        Active substance according to the invention                                                          50.0      mg                                           Lactose                250.0     mg                                           Suppository mass q.s. ad                                                                             1.7       g                                            ______________________________________                                    

Preparation

The active substance and lactose are mixed together and the mixture isuniformly suspended in the molten suppository mass. The suspensions arepoured into chilled moulds to form suppositories weighing 1.7 g.

    ______________________________________                                        g) Ampoules                                                                   ______________________________________                                        Active substance according to the invention                                                          20.0      mg                                           Sodium chloride        5.0       mg                                           Twice distilled water q.s. ad                                                                        2.0       ml                                           ______________________________________                                    

Preparation

The active substance and the sodium chloride are dissolved in twicedistilled water and the solution is transferred under sterile conditionsinto ampoules.

    ______________________________________                                        h) Ampoules                                                                   ______________________________________                                        Active substance according to the invention                                                          10.0      mg                                           Sodium chloride        7.0       mg                                           Twice distilled water q.s. ad                                                                        1.0       ml                                           ______________________________________                                    

    ______________________________________                                        i) Drops                                                                      ______________________________________                                        Active substance according to the invention                                                          0.70      g                                            Methyl p-hydroxybenzoate                                                                             0.07      g                                            Propyl p-hydroxybenzoate                                                                             9.03      g                                            Demineralised water q.s. ad                                                                          100.00    ml                                           ______________________________________                                    

Preparation

The active substance and preservatives are dissolved in demineralisedwater, the solution is filtered and transferred into 100 ml vials.

What is claimed is:
 1. A compound of formula I ##STR183## wherein: Arepresents benzo;m represents 2 or 3; R¹ represents thienyl ortert-butyl; each R² independently represents; R³ and R⁴ independentlyrepresent(a) hydrogen, or (b) branched or unbranched C₃₋₆ alkenyl, or(c) branched or unbranched C₃₋₆ alkynyl, or (d) branched or unbranchedC₁₋₁₂ alkyl which may be optionally mono- or Di-substituted by:hydroxy,or (C₁₋₄) alkoxy, or Di(C₁₋₄)alkoxy, or Di (C₁₋₄) alkylamino, or furyl,or pyridyl, or pyrrolidinyl or N-methylpyrrolidinyl, or morpholinyl, orindolyl, or nitrilo, or thienyl, or adamantyl, or cyclohexyl, ornaphthyloxy, or phenoxy or phenyl wherein the phenyl group may beoptionally mono, Di- or trisubstituted by hydroxy, (C₁₋₄)alkyl,(C₁₋₄)alkoxy, benzyloxy, F, Cl, Br, I, CF₃, N₃, adamantyl, --SO₂ NH₂,NHCOCH₃, or by the bridge --O--CH₂ --O; or R³ represents hydrogen and R⁴represents phenyl, fluorophenyl, cyclohexyl, pyridyl or N-benzylpyridyl;or R³ and R⁴ together with the nitrogen atom to which they are bondedrepresent pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl orpiperazinyl, whilst the piperazinyl ring may optionally be N-substitutedby methyl, unsubstituted phenyl, mono- or Di(C₁₋₄) alkoxyphenyl,cyano-substituted phenyl, pyrimidinyl, phenyl (C₁₋₄)alkyl,(C₁₋₄)alkylphenyl or ##STR184## or the salts thereof withphysiologically acceptable acids.
 2. A compound having the formula:##STR185## or the salts thereof with physiologically acceptable acids orcomplex-forming agents.
 3. A pharmaceutical composition comprising acompound according to claim 1, or 2 and a pharmaceutically acceptablecarrier.
 4. A compound of formula I ##STR186## wherein: A representsbenzo;m represents 2 or 3; R¹ represents thienyl; each R² independentlyrepresents hydroxy, C₁₋₄ alkoxy, benzyloxy, F, Cl, Br, I, C₁₋₄ alkyl,methanesulfonyloxy or methanesulfonamido, or two adjacent substituents;R² may represent --O--CH₂ --O-- or --O--CH₂ --CH₂ --O--; R³ and R⁴independently represent(a) hydrogen, or (b) branched or unbranched C₃₋₆alkenyl, or (c) branched or unbranched C₃₋₆ alkynyl, or (d) branched orunbranched C₁₋₁₂ alkyl which may be optionally mono- or Di-substitutedby:hydroxy, or (C₁₋₄) alkoxy, or Di(C₁₋₄)alkoxy, or Di(C₁₋₄)alkylamino,or furyl, or pyridyl, or pyrrolidinyl or N-methylpyrrolidinyl, ormorpholinyl, or indolyl, or nitrilo, or thienyl, or adamantyl, orcyclohexyl, or naphthyloxy, or phenoxy or phenyl wherein the phenylgroup may be optionally mono, Di- or trisubstituted by hydroxy,(C₁₋₄)alkyl, (C₁₋₄)alkoxy, benzyloxy, F, Cl, Br, I, CF₃, N₃, adamantyl,--SO₂ NH₂, NHCOCH₃, or by the bridge --O--CH₂ --O; or R³ representshydrogen and R⁴ represents phenyl, fluorophenyl, cyclohexyl, pyridyl orN-benzylpyridyl; or R³ and R⁴ together with the nitrogen atom to whichthey are bonded represent pyrrolidinyl, piperidinyl, morpholinyl,thiomorpholinyl or piperazinyl, whilst the piperazinyl ring mayoptionally be N-substituted by methyl, unsubstituted phenyl, mono- orDi(C₁₋₄)alkoxyphenyl, cyano-substituted phenyl, pyrimidinyl,phenyl(C₁₋₄)alkyl, (C₁₋₄)alkylphenyl or ##STR187## or the salts thereofwith physiologically acceptable acids.
 5. The compound according toclaim 4, wherein m is 2 and each R² is independently methoxy, hydroxy,benzyloxy, methyl or chlorine or together are --OCH2-- when R² is inposition 6 and
 7. 6. A pharmaceutical composition comprising a compoundaccording to claim 4 and a pharmaceutically acceptable carrier.
 7. Amethod for providing protection against stroke in a warm-blooded animalwhich comprises administering to said animal a therapeutically effectiveamount of a pharmaceutical composition according to claim 3 or 6.